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Research Reagents

Cas9 protein/gRNA transfection reagent
GenomONE TM-GE EX

Product name Cat. # Specification Frequency of use
(96-well plate)
Freeze-dried HVJ-E Reagent F,G Buffer
GenomONE-GE EX GG001EX 1 vial each 1 vial 1 vial 325
GG004EX 4 vials each 1 vial 1 vial 1,300
GG016EX 16 vials each 4 vials 2 vials 5,200

GenomONE-GE EX enables efficient Cas9 protein/gRNA transfection in a variety of difficult-to-transfect cells.
In case of plasmid DNA transfection for Cas9 and gRNA expression, please use GenomONE-GX EX .

*This product is sold for research purpose only. It may not be used for treatment or other clinical purposes or for intra- and extracorporeal diagnosis in humans or animals.

Features of GenomONE-GE EX

  • Applicable for CRISPR-Cas9-mediated knock-out and knock-in
  • Applicable for difficult-to-transfect cells (e.g., Mouse primary T cell, U-937, Jurkat)
  • Simple operability (completion of transfection within 10 min)

Efficient delivery of Cas9 protein and gRNA into mouse primary T cells

T cells isolated from splenocytes were transfected with Cas9 protein sgRNA (2’OMe and PS modifications) using HVJ-E (GenomONE-GE EX) and product C or T (competitors). Genome editing efficiency was evaluated using T7 endonuclease I mismatch cleavage assay two days after transfection.

Induction of cell death by delivery of Cas9 protein and lethal gRNA targeting thousands of locations in the genome

Cas9 protein and Edit-R Lethal control gRNA (Horizon Discovery) were delivered into mouse primary T cells and Jurkat cells using GenomONE-GE EX and product C or T (competitors). Cell viability was measured by wst-8 assay (A450) two days after transfection. The GenomONE-GE EX efficiently delivered Cas9 protein and gRNA into mouse primary T cells and Jurkat cells, which are known as difficult-to-transfect cells.

Highly efficient ssODN-mediated homology directed repair (HDR)

Cas9 protein, sgRNA and ssODN (donor DNA) were transfected into HeLa, A549, HL-60 and U-937 cells using GenomONE-GE EX, respectively. ssODN (66base) was designed to carry the recognition site of BamHI restriction enzyme. After transfection, NU 7441(Tocris), an inhibitor of DNA-PK which blocks non-homologous end joining (NHEJ), was added to cells for 24 hours. Knock-in efficiency was evaluated using restriction fragment length polymorphism (RFLP) two days after transfection.

Delivery of Cas9 protein and gRNA into different cells

Cas9 protein and gRNA (2-part gRNA or sgRNA) were delivered into HeLa, RAW264.7, Jurkat, THP-1, K-562, and Raji cells using GenomONE-GE EX. Genome editing efficiency was evaluated using T7 endonuclease I mismatch cleavage assay two days after transfection.

Improvement of genome editing efficiency using chemically modified sgRNA

Unmodified or 2’OMe+PS-modified sgRNA was delivered with Cas9 protein into U-937 cells, respectively. Two days after transfection, genome editing efficiency was evaluated using T7 endonuclease I mismatch cleavage assay.

Improvement of cytotoxicity using detached floating cells in the transfection

HEK-293 cells were trypsinized, neutralized, centrifuged, and re-suspended in 10% fresh FBS culture medium. The detached floating cells were transfected with Cas9 protein and PPIB-targeting sgRNA using HVJ-E (GenomONE-GE EX ); centrifuged, re-resupended in 10% fresh FBS culture medium, and then transferred to 48-well culture plate. Genome editing efficiency was evaluated using T7 endonuclease I mismatch cleavage assay two days after transfection.
If severe cytotoxicity and/or many fused cells are observed in transfection of adherent cells such as HEK-293 cells, it is possible to reduce cytotoxicity and/or cell-to-cell fusion using detached floating cells in the transfection as described above.

Related technical information

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